Remote Control of Bovine Catalase Activity with Specific Frequencies of Human and Bovine Catalase with and without Bound NADP+

Remote control enzyme technology is widely used today through resonance. In this study, we showed that the use of frequencies of the catalase enzyme itself to increase enzymatic rate is successful not only in test tubes but also remotely. The present study also suggests that, under optimal temperature, the use of bovine catalase frequency (the specific frequency of that enzyme) has a superior rate promoting vibration than the human catalase frequency, and so increases very significantly the chemical rate of bovine catalase (about 120% at 40˚C). It also suggests that bovine catalase subjected to bovine and human frequencies with catalase bound NADP+ experienced more resonance weight towards NADP+ and so were more slowly reduced back to catalase bound NADPH, increasing compound II formation rate, and slowing down the catalase activity rate.

细叶百合Catalase基因的克隆及表达分析

为探究细叶百合(Lilium pumilum)Catalase(CAT)基因与耐盐碱胁迫的关系,从细叶百合鳞茎中成功克隆出LpCat基因。该基因的ORF区长度为1479 bp,编码492个氨基酸,对其进行序列比对和系统进化树分析,发现细叶百合Catalase蛋白与通江百合(L.sargentiae)、菠萝(Ananas comosus)、油棕(Elaeis guineensis)等植物的Catalase蛋白亲缘关系较近。构建了pQE-LpCat原核表达载体,以1 mol·L^(-1)IPTG为诱导剂进行LpCat蛋白的体外诱导表达与纯化。在添加50 mmol·L^(-1)NaHCO_(3)胁迫下,携带PQE-LpCat蛋白的菌液生长浓度高于对照菌株。在1 mol·L^(-1)NaHCO_(3)、2.5 mol·L^(-1)H_(2)O_(2)胁迫下过表达LpCAT基因烟草(Nicotiana plumbaginifolia)植株与野生型相比,枯萎程度相对较低。通过净光合速率、气孔导度、胞间CO_(2)和蒸腾速率,H_(2)O_(2)含量,丙二醛(MDA)等生理指标检测说明过表达LpCat基因烟草植株比野生型烟草植株更耐盐碱。

Effects of Hg^(2+) on Isozymes of Peroxidase,Catalase and Superoxide Dismutase in Wheat Seedlings

[Objective] This work was aimed to explore the mechanism of Hg2+ toxicity on plants.[Method]Activities of peroxidase(POD),catalase(CAT)and superoxide dismutase(SOD)were investigated in wheat(Triticum aestivum L.)seedlings under Hg2+ stress at different concentrations.[Result]① There were no obvious effects on the growth of seedlings when the concentration of Hg2+ was lower than 0.10 mmol/L.However,toxic effects on the growth of seedling were observed when the concentration of Hg2+ was higher than 0.10 mmol/L.② Different tissues showed different resistant ability in response to Hg2+ stress.The leaves and roots of wheat seedlings were more insensitive to Hg2+ toxicity.③ CAT was more sensitive to Hg2+ stress compared to POD and SOD.[Conclusion]The toxic effect was related to the concentration of Hg2+(0.10 mmol/L).The higher concentration of Hg2+ could affect the expression of POD,CAT,and SOD isozymes in the leaves,roots of wheat seedlings and germinated seeds,which further affect the normal metabolism of membrane lipid and inhibit the growth of wheat seedlings at last.

幽门螺杆菌Catalase基因的克隆、表达及其抗原性鉴定

目的：构建含人幽门螺杆菌（Hpylori，Hp）过氧化氢酶（catalase，KatA）编码基因的重组质粒，测定、分析其核酸序列，并在E.coli中表达，研究其抗原性。方法：应用PCR技术从HpDNA染色体中扩增KatA编码基因片段，将其T-A克隆和测序，并与GenBank公布的其他Hp菌株的基因序列比较，再将目的基因插入至融合表达载体pGEX-4T-1中进行表达，用GST亲和层析对其进行纯化。纯化产物用于对29株小鼠抗Hp-全菌单克隆抗体（mAb）的鉴定及与Hp啊感染患者血清进行Western blot。结果：KatA基因全长为1 515 bp，并在GenBank上登录（No．DQ333889），与GenBank公布的其他Hp菌株的核酸的同源性为96％～97％，表达的KatA融合蛋白的相对分子质量（Mr）为85 000，29株小鼠抗Hp全菌mAb中有4株mAb是针对KatA的，表达产物可被Hp感染患者的血清特异性识别。结论：重组KatA具有较好的抗原性，为坳检测试剂和疫苗的研究奠定了基础。

Cloning of catalase and expression patterns of catalase and selenium-dependent glutathione peroxidase from Exopalaemon carinicauda in response to low salinity stress

Catalase （CAT） and selenium-dependent glutathione peroxidase （Se-GPx） play a vital role in protecting organisms against various oxidative stresses by eliminating H202, The objective of this paper is to evaluate the roles of these antioxidant molecules in the ridgetail white prawn Exopalaemon carinicauda in response to low salinity stress. A complementary DNA （cDNA） containing the complete coding sequence of CAT was cloned from the hepatopancreas using reverse-transcription polymerase chain reaction （RT-PCR） and rapid amplification of cDNA ends. The full-length cDNA of CAT （2 649 bp） contains a 5＇-untranslated region （UTR） of 78 bp, a 3＇- UTR of 1 017 bp, with a poly （A） tail, and an open reading frame of 1 554 bp encoding a 517-amino-acid polypeptide with predicted molecular mass of 58.46 kDa and estimated isoelectric point of 6.64. This CAT sequence contained the proximal active site signature （60FDRERIPERWHAKGAG76）, proximal heme-ligand signature sequence （350RLFSYPDTH358） and three catalytic amino acid residues （His71, Asn144 and Tyr354）. Sequence comparison showed that the CAT deduced amino acid sequence of E. carinicauda shared 68%-92% of identities with those of other species. Quantitative real-time PCR analysis revealed that CAT mRNA was widely expressed in the hepatopancreas （highest）, hemocyte, eyestalk, heart, gill, muscle, ovary and stomach. Under low salinity stress, CAT and GPx mRNA expression levels both in the gill and hepatopancreas increased significantly at the first 48 h and 6 h respectively, indicating a tissue- and time-dependent antioxidant response in E. carinicauda. All these results indicate that E. carinicauda CAT is a member of the CAT family and might be involved in the acute response against low salinity stress.

幽门杆菌Catalase/GST融合蛋白的表达、标签切除及鉴定

旨在利用GST融合基因表达系统表达幽门螺杆菌Catalase融合蛋白,并利用凝血酶切除GST标签。将重组质粒Catalase/pGEX-4T-1转化大肠杆菌BL21(DE3)感受态中,用IPTG进行诱导表达,菌体经反复冻融、溶菌酶裂解及超声破菌后,Catalase/GST融合蛋白以部分可溶性的形式表达在上清中。采用谷胱甘肽琼脂糖树脂Glutathione Sepharose 4B对其进行纯化,得到Catalase/GST融合蛋白,再用凝血酶进行GST标签的切除,所得产物进行Western blotting鉴定。高效表达出Catalase/GST融合蛋白的相对分子质量约85 kD,凝血酶成功地切除了GST标签,Western blotting证实Catalase蛋白能被鼠抗Catalase单克隆抗体识别。

Effects of Catalase and Ascorbate Peroxidase on the Root Growth of Rice under Cadmium Stress

[Objective] The research aimed to study the correlations between catalase（CAT） and ascorbate peroxidase（ASP） and the growth and development of rice roots under cadmium stress.[Method] Taking rice variety Zhonghua No.11 as materials,the changes of rice seedlings under the treatment conditions of Cd,Cd＋CAT inhibitor,Cd＋APX inhibitor were studied.[Result] Under Cd stress,inhibition of CAT activity caused the significant inhibition on the growth of aerial parts,decreased the number of adventitious roots and lateral roots,but it can significant promote the elongation growth of adventitious roots and lateral roots.Moreover,the length of the first lateral root from root tip on the primary roots and adventitious roots was also increased than control.When APX activity was inhibited,the growth changes of rice were similar with that treated by CAT inhibitor.[Conclusion] CAT and APX may play important roles in the regulation of rice root system growth in both non-stress and Cd-stressed rice

Cloning and Differential Gene Expression of Two Catalases in Suaeda salsa in Response to Salt Stress

Two different cDNA clones (Sscat1 and Sscat2) encoding catalase, the primary important H2O2-scavenging enzyme, were isolated from a AZap-cDNA library constructed from a 400 mmol/L NaCl-treated library of Suaeda salsa ( L.) Pall aerial tissue. Sscat1 (1.7 kb) contains a full open reading frame of 492 amino acids and Sscat2 (1.1 kb) is a partial clone. BLAST analysis indicates that the two clones share 71.9% identity in nucleotide sequence and 75% identity in deduced amino acid sequence within the last 287 amino acid residues of Sscat1. Southern blotting analysis showed that Sscat1 is multicopy in S. salsa genome, while Sscat2 is a single copy gene. Northern blotting analysis showed a rapid increase in the steady-level of both genes in roots after 48 It salt treatment, but only Sscat1 was induced in salinity treated leaves. Time-course analysis carried out in leaves confirmed that Sscat1 was induced by salt stress, in contrast to Sscat2. These implied that the expression of Sscat1 and Sscat2 genes are differentially regulated in S. salsa. The activity of total catalase is dramatically increased in response to salt stress.

Association of Catalase Genotype with Oxidative Stress in the Predication of Colorectal Cancer:Modification by Epidemiological Factors

Objective This paper aims to assess the interaction between common variations in catalase（CAT） polymorphic gene and environmental factors for antioxidant defense enzyme in modulating individual susceptibility to colorectal cancer（CRC）.Methods A case-control study with 880 colorectal cancer cases and 848 controls was conducted to investigate whether variations in the catalase（CAT） gene,one of the genes involved in scavenging oxidative stress,influenced susceptibility to CRC.Results The interaction between life style and genotypes as well as with their effects on colorectal cancer was deduced from the present study.Significant difference（P=0.01） was identified in the distribution of CAT genotype between the colorectal cancer cases and the controls.The CRC cases had significantly lower mean activity than the controls（P〈0.01）.Correlation analyses revealed statistically significant correlations between CAT activity and CAT genotype（P〈0.01）.Conclusion The risk of CRC was associated with smoking,low vegetable consumption,high pork and poultry consumptions,and low or high BMI.This is the first study reporting an association of polymorphism CAT-21A〉T with colorectal cancer.Low CAT activity was associated with an increased risk of CRC;however,no evidence was found to support an association between CAT-21A〉T polymorphism and CRC risk.

玉米catalase-3基因克隆及低温表达研究

应用RT-PCR技术,从玉米自交系承18经14℃冷锻炼处理的芽鞘中扩增出长1491 bp的Cat3全长cDNA。以pGEM-T Easy为载体,克隆了扩增片段,经酶切鉴定和DNA序列测定,确认为玉米Cat3基因。序列分析结果表明,该序列与GenBank中accession number:L05934的序列仅在两个核苷酸位点存在差异。同时采用Northern杂交方法对Cat3在不同低温处理的玉米芽鞘中的表达特性进行了研究,结果显示Cat3在经14℃冷锻炼处理的芽鞘中表达增强,表明该基因受低温的诱导表达,为玉米抗寒基因,对提高玉米的抗寒力起积极作用。

丝瓜过氧化氢酶(Catalase,CAT)酶学特性的研究

为研究丝瓜过氧化氢酶(Catalase,CAT)酶学特性,本研究应用分光光度计开展温度、pH、酶体积、反应底物浓度等对酶反应的影响,为丝瓜酶促褐变的研究提供基础。结果表明,CAT最适反应体系:以H_2O_2为底物,最适底物浓度为0.33 mol/L H_2O_2,Km值为0.1818 mol/L,最适温度为40℃,最适pH值为7.8,最适酶液体积为0.1 ml。

Catalase活性在PTL诱导MM凋亡不同敏感性中的作用研究

目的研究小白菊内酯(PTL)对多发性骨髓瘤细胞敏感性的不同及机制,为PTL的联合化疗策略提供理论依据。方法以多发性骨髓瘤细胞KMM-1、MM1S、NCI-H929、KMS-5为研究对象,利用DAPI检测细胞凋亡、Weston blot检测过氧化氢酶(Catalase)蛋白的表达、流式细胞仪检测活性氧水平、Catalase-520和GPX-340试剂盒检测Catalase和谷胱甘肽过氧化物酶活性。结果KMS-5、NCI-H929与KMM-1、MM1S对2.5μmol/L、5μmol/L PTL凋亡率具有显著差异性(P<0.01)。在PTL 2.5μmol/L时活性氧增高与药物作用时间关系中,KMM-1、MM1S与KMS-5、NCI-H929相比,峰值显著升高(P<0.01)。在MM1S和KMM-1细胞Catalase的表达水平、活性与NCI-H929,KMS-5相比明显降低,Catalase抑制剂3-氨基-1,2,4-三唑提高NCI-H929,KMS-5对PTL介导的细胞凋亡。结论Catalase活性是PTL敏感性的关键因素,与Catalase抑制剂药物合用可以提高PTL更有效的抗MM的作用。

苯系物对仿刺参catalase基因表达及酶活性的影响

以仿刺参（Apostichopus japonicus）为受试生物,采用半静水式试验方法,设置3种不同浓度（1/5、1/25、1/125的96 h-LC50）的苯、甲苯、乙基苯、邻–二甲苯、间–二甲苯和对–二甲苯处理健康仿刺参,检测仿刺参过氧化氢酶（CAT）基因在呼吸树、肠组织中的表达和酶活性变化情况。结果发现：在各苯系物处理组的仿刺参呼吸树和肠组织中,cat基因的转录表达变化显著;苯、甲苯、乙基苯、邻–二甲苯对呼吸树中CAT活性具有诱导作用,其中乙基苯的诱导倍数最高,为12.0~19.8倍;6种苯系物对肠组织中CAT活性具有抑制作用,抑制程度大小顺序为：邻–二甲苯〉乙基苯〉对–二甲苯〉甲苯〉间–二甲苯〉苯。表明苯系物对仿刺参呼吸树、肠具有氧化胁迫作用,可能造成2种组织的氧化损伤。相关性分析表明：苯系物处理后,仿刺参肠组织中cat m RNA相对表达倍数与CAT活性变化呈显著正相关;仿刺参肠和呼吸树中cat m RNA相对表达倍数变化呈显著正相关。以上结果为苯系物对仿刺参的生物毒性评价提供了基础数据。

Response of soil catalase activity to chromium contamination

The impact of chromium（Ⅲ） and （Ⅵ） forms on soil catalase activity was presented. The Orthic Podzol, Haplic Phaeozem and Mollic Gleysol from different depths were used in the experiment. The soil samples were amended with solution of Cr（Ⅲ） using CrCl3, and with Cr（Ⅵ） using K2Cr2O7 in the concentration range from 0 to 20 mg/kg, whereas the samples without the addition of chromium served as control. Catalase activity was assayed by one of the commonly used spectrophotometric methods. As it was demonstrated in the experiment, both Cr（Ⅲ） and Cr（Ⅵ） have an ability to reduce soil catalase activity. A chromium dosage of 20 mg/kg caused the inhibition of catalase activity and the corresponding contamination levels ranged from 75% to 92% for Cr（Ⅲ） and 68% to 76% for Cr（Ⅵ）, with relation to the control. Catalase activity reached maximum in the soil material from surface layers （0-25 cm）, typically characterized by the highest content of organic matter creating favorable conditions for microorganisms.

基于Cat.1技术与阵列式位移计的深基坑监测方法与应用

深基坑开挖的危险系数高,容易引发安全事故,需要采用有效的地质灾害监测系统。为实现深基坑施工阶段变形的有效监测与预警,提出了采用基于Cat.1技术与阵列式位移计的无线自动化监测方法。首先通过与传统的LoRa通信技术对比,分析了Cat.1技术的特点和优势。然后把Cat.1技术与阵列式位移计共同应用于基坑监测,建立了监测系统。最后,以某深基坑监测项目为例,分析了代表性测点的水平位移监测结果。结果表明,所建立的监测系统能够稳定地采集监测数据并上传至监测平台,实时反映基坑测点水平位移变化规律,且监测结果的可解释性高,有助于及时发出预警信号,做好风险防范措施。

槟榔Catalase 基因的克隆及亚细胞定位

过氧化氢酶(Catalase)广泛存在于生物体内,主要功能是清除植物细胞内光呼吸、线粒体电子传递及脂肪β-氧化等过程中产生的H2O2,从而保护细胞免于过氧化氢酶的毒害。为了解槟榔过氧化氢酶基因的相关信息,利用植物总RNA提取试剂盒提取槟榔叶片总RNA,通过同源克隆和RACE技术获得槟榔全长cDNA,并对其进行生物信息学分析及亚细胞定位。结果表明:获得ArCAT1(MN692600)、ArCAT2(MN692601)、ArCAT3(MN692602)3个同源基因,其序列全长均为1476 bp,编码492个氨基酸。遗传进化分析显示,ArCATS与同为棕榈科的油棕(Elaeis guineensis)、海枣(Phoenix dactylifera)等植物的过氧化氢酶氨基酸序列具有较高的相似性,其中与油棕的亲缘关系最近。亚细胞定位结果表明,ArCAT1蛋白定位于过氧化物酶体中,而ArCAT2和ArCAT3既定位于过氧化物酶体又定位于细胞核中。这为进一步研究Catalase基因在槟榔中的生物学功能奠定基础。

Modulatory effect of pineapple peel extract on lipid peroxidation,catalase activity and hepatic biomarker levels in blood plasma of alcoholinduced oxidative stressed rats

Objective:To investigate the ability of the methanolic extract of pineapple peel to modulate alcohol-induced lipid peroxidation,changes in catalase activities and hepatic biochemical marker levels in blood plasma.Methods:Oxidative stress was induced by oral administration of ethanol(20%w/v) at a dosage of 5 niL/kg bw in rats.After 28 days of treatment,the rats were fasted overnight and sacrificed by cervical dislocation.Blood was collected with a 2 mL syringe by cardiac puncture and was centrifuged at 3000 rpm for 10 min.The plasma was analyzed to evaluate malondialdehyde(MDA),catalase activity,aspartate aminotransferase(AST),alkaline phosphatase(ALP) and alanine aminotransferase(ALT) concentrations.Results:Administration of alcohol caused a drastic increase(87.74%) in MDA level compared with the control.Pineapple peel extract significantly reduced the MDA level by 60.16%at 2.S mL/kg bw.Rats fed alcohol only had the highest catalase activity,treatment with pineapple peel extract at 2.5 mL/kg bw however, reduced the activity.Increased AST,ALP and ALT activities were observed in rats fed alcohol only respectively,treatment with pineapple peel extract drastically reduced their activities. Conclusions:The positive modulation of lipid peroxidation,catalase activities as well as hepatic biomarker levels of blood plasma by the methanolic extract of pineapple peels under alcoholinduced oxidative stress is an indication of its protective ability in the management of alcoholinduced toxicity.

Response of Superoxide Dismutase,Catalase,and ATPase Activity in Bacteria Exposed to Acetamiprid

Objective To investigate how acetamiprid, a new insecticide, affects the activity of superoxide dismutase （SOD）, catalase （CAT）, and ATPase and the SOD isozyme patterns in two G-bacteria, E. coli K12 anti Pse.FH2, and one G^＋ bactemm, B. subtilis. Methods The SOD, CAT, and ATPase specific activities of cell lysates were determined spectrophotometrically at 550 nm, 240 nm, and 660 nm, respectively, with kits A001, A016, and A007. SOD isozyme patterns were detected by native PAGE analysis. Results SOD and CAT activities in the tested bacteria increased significantly in a concentration-dependent manner after different concentrations of acetamiprid were applied. The activity of SOD in B. subtilis and Pse.FH2 was stimulated and reached the highest level after treatment with 100 mg/L acetamiprid for 0.5 h. For Pse.FH2, there was another stimulation of SOD activity after acetamiprid application for about 8.0 h and the second stimulation was stronger than the first. The stimulation by acetamiprid showed a relative lag for E. coli K12. Acetamiprid seemed to exhibit a similar effect on CAT activity of the two G bacteria and had an evident influence on ATPase activity in the three bacteria within a relatively short period. Only one SOD isozyme was detectable in Pse.FH2 and B. subtilis, while different isozyme compositions in E. coli could be detected by native PAGE analysis. Conclusion Acetamiprid causes a certain oxidative stress on the three bacteria which may not only elevate SOD and CAT activities but also generate new SOD isozymes to antagonize oxidative stress. However, this oxidative stress lasts for a relatively short time and does not cause a long-term damage.

Molecular cloning, characterization and expression analysis of a catalase gene in Paphia textile

Catalase is an important antioxidant protein that can protect organisms against various forms of oxidative damage by eliminating hydrogen peroxide. In this study, the catalase c DNA of Paphia textile（Pt CAT） was cloned using RTPCR and rapid amplification of c DNA ends（RACE）. Pt CAT is 1 921 bp long and consists of a 5′-UTR of 50 bp, a 3′-UTR of 349 bp, and an ORF of 1 542 bp that encodes 513 amino acids with a molecular weight of 58.4 k D and an estimated isoelectric point of 8.2. Sequence alignment indicated that Pt CAT contained a highly conserved catalytic signature motif（^（61）FNRERIPERVVHAKGAG^（77））, a proximal heme-ligand signature sequence（^（352）RLFSYSDP^（359））, and three catalytic amino acid residues（H^（72）, N^（145）, and Y^（356））. Pt CAT also contains two putative N-glycosylation sites（^（34）NKT^（36） and ^（437）NFT^（439）） and a peroxisome-targeting signal（^（511）AQL^（513））. Furthermore, Pt CAT shares 53%–88% identity and 29%–89% similarity with other catalase amino acid sequences. Pt CAT m RNA was present in all tested organs, including the heart, digestive gland, adductor muscle, gonad, gill, and mantle, but its expression was highest in the digestive gland. High-temperature-induced stress produced two expression patterns of Pt CAT m RNA： first, an initial up-regulation followed by a down-regulation in the heart, digestive gland, and gonad and, second, consistent down-regulation in all other organs. These results demonstrate that Pt CAT is a typical member of the catalase family and might be involved in the responses to harmful environmental factors.

HIGHLY SENSITIVE CATALASE ELECTRODE BASED ON POLYPYRROLE FILMS WITH MICROCONTAINERS

Highly sensitive catalase electrodes for sensing hydrogen peroxide have been fabricated based on polypyrrole films with microcontainers. The microcontainers have a cup-like morphology and are arranged in a density of 4000 units cm^-2. Catalase was immobilized into the polypyrrole films with microcontainers （Ppy-mc）, which were coated on a Pt substrate electrode. The catalase/Ppy-mc/Pt electrode showed linear response to hydrogen peroxide in the range of 0-18 mmol/L at a potential of-0.3 V （versus SCE）. Its sensitivity was measured to be approximately 3.64 μA （mmol/L） ^-1 cm^- 2, which is about two times that of the electrode fabricated from a flat Ppy film （catalase/Ppy-flat/Pt electrode）. The electrode is highly selective for hydrogen peroxide and its sensitivity is interfered by potential interferents such as ascorbic acid, urea and fructose. Furthermore, such catalase electrodes showed long-term storage stability of 15 days under dry conditions at 4℃.